Abstract：By means of a new method,gel-ligation method,the recombinant expression plasmid carrying genes coding to Schistosoma japonicum antigen epitope peptide was rapidly and efficiently constructed.In this method,the plasmid pUMCV was digested with restriction endonucleases Sal I and EcoRI,and the digested products were subjected to electrophoresis in low-melting point agarose gel.Then,the digested plasmid in gel was sliced under ultra-violet lamp.Single or multiple gene cDNA coding to antigen epitope peptide was ligated together,and the ligated product was directly inserted into digested plasmid in the presence of low-melting point agarose gel.The recombinant plasmid was then transformed to E.coli DH5α strain,and the colonies were picked out and characterized by restriction endonuclease digestion and sequence analysis.It was demonstrated that the cDNAs coding to S.japonicum antigen epitope peptide could be inserted successively into vector plasmid.It is evident that the gel-ligation method is proved to be a simple and efficient method to make recombinant constructs,which would provide a new tool for the rapid and convenient cloning and further screening of vaccine candidate epitope of S.japonicum.
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